f78b8c4c7e8c23acd998ce19c7ef5af7c404adc4 angie Wed Dec 2 16:54:28 2020 -0800 Use Yatish's find_parsimonious_assignments program to make parsimony score bigWigs. Also save full output of find_parsimonious_assignments for collaborators (Russ request). diff --git src/hg/utils/otto/sarscov2phylo/processRelease.sh src/hg/utils/otto/sarscov2phylo/processRelease.sh index 22b7fe6..979fcc4 100755 --- src/hg/utils/otto/sarscov2phylo/processRelease.sh +++ src/hg/utils/otto/sarscov2phylo/processRelease.sh @@ -1,123 +1,148 @@ #!/bin/bash set -beEu -x -o pipefail # Do not modify this script, modify the source tree copy: # kent/src/hg/utils/otto/sarscov2phylo/processRelease.sh usage() { echo "usage: $0 releaseLabel problematic_sites_sarsCov2.vcf" } if [ $# != 2 ]; then usage exit 1 fi releaseLabel=$1 problematicSitesVcf=$2 echo "releaseLabel=$releaseLabel problematicSitesVcf=$problematicSitesVcf" scriptDir=$(dirname "${BASH_SOURCE[0]}") # UShER (and matToVcf) usherDir=~angie/github/usher usher=$usherDir/build/usher matToVcf=$usherDir/build/matToVcf +# strain_phylogenetics / find_parsimonious_assignments for parsimony scores +find_parsimonious_assignments=~angie/github/strain_phylogenetics/build/find_parsimonious_assignments + # Files generated by getRelease.sh metadata=gisaid_metadata.tsv fasta=gisaid_sequences.fasta alignedFasta=gisaid_aligned.fasta # We'll need to prune the tree to keep only those sequences that are in metadata and fasta: grep ^\> $fasta | sed -re 's/^>//' | sort > namesInFasta # Make renaming files for tree (EPI_ IDs) and VCF (isolate names): add strain name and # date with shortened year. awk -F"\t" '{print $3 "\t" $1 "|" $3 "|" $5;}' $metadata \ | grep -Fwf namesInFasta \ | sed -re 's/\|20/\|/;' \ | sort > tree.renaming awk -F"\t" '{print $1 "\t" $1 "|" $3 "|" $5;}' $metadata \ | grep -Fwf namesInFasta \ | sed -re 's/\|20/\|/;' \ | sort > vcf.renaming wc -l tree.renaming wc -l vcf.renaming # Rename tree IDs from just EPI IDs to strain|epi|date ids and prune any sequences not in # metadata and fasta: phyloRenameAndPrune ft_SH.tree tree.renaming ft_SH.renamed.tree sed -re 's/\)[0-9.]+:/\):/g; s/:[0-9e.:-]+//g; s/[\(\);]//g; s/,/\n/g;'" s/'//g;" \ ft_SH.renamed.tree \ | sort > tree.renamedPruned.ids.sorted wc -l tree.renamedPruned.ids.sorted # Set the root of the tree to the Wuhan/Hu-1 (NC_045512.2) reference not WIV04: ~angie/github/newick_utils/src/nw_reroot ft_SH.renamed.tree \ 'Wuhan/Hu-1/2019|EPI_ISL_402125|19-12-26' > ft_SH.reroot.nh # Run faToVcf without any filtering, masking or conversion of ambiguous alleles: time faToVcf $alignedFasta stdout -includeRef \ -ref=Wuhan/Hu-1/2019 \ -vcfChrom=NC_045512v2 -verbose=2 \ | vcfRenameAndPrune stdin vcf.renaming gisaid-$releaseLabel.unfiltered.vcf ls -l gisaid-$releaseLabel.unfiltered.vcf wc -l gisaid-$releaseLabel.unfiltered.vcf gzip -f gisaid-$releaseLabel.unfiltered.vcf # Run usher and matToVcf to make ambig-resolved VCF for hgTracks time $usher -c \ --vcf gisaid-$releaseLabel.unfiltered.vcf.gz \ --tree ft_SH.reroot.nh \ --save-mutation-annotated-tree sarscov2phylo-$releaseLabel.notMasked.pb \ --write-uncondensed-final-tree mv uncondensed-final-tree.nh ft_SH.reroot.collapsed.notMasked.nh $matToVcf -i sarscov2phylo-$releaseLabel.notMasked.pb \ -v gisaid-$releaseLabel.vcf bgzip -f gisaid-$releaseLabel.vcf tabix -p vcf gisaid-$releaseLabel.vcf.gz # Remove problematic sites recommended for masking for usher/phyloPlace tawk '{ if ($1 ~ /^#/) { print; } else if ($7 == "mask") { $1 = "NC_045512v2"; print; } }' \ $problematicSitesVcf > mask.vcf time vcfFilter -excludeVcf=mask.vcf gisaid-$releaseLabel.unfiltered.vcf.gz \ | gzip -c \ > gisaid-$releaseLabel.unfiltered.masked.vcf.gz +# Full find_parsimonious_assignments output on uncollapsed tree for collaborators +time $find_parsimonious_assignments --tree ft_SH.reroot.nh \ + --vcf gisaid-$releaseLabel.unfiltered.masked.vcf.gz \ +| gzip -c \ + > find_parsimonious_assignments.$releaseLabel.out.gz + # Run usher to collapse tree and make masked protobuf for hgPhyloPlace time $usher -c \ --vcf gisaid-$releaseLabel.unfiltered.masked.vcf.gz \ --tree ft_SH.reroot.nh \ --save-mutation-annotated-tree sarscov2phylo-$releaseLabel.masked.pb \ --write-uncondensed-final-tree mv uncondensed-final-tree.nh ft_SH.reroot.collapsed.nh +# Parsimony scores on collapsed tree +time $find_parsimonious_assignments --tree ft_SH.reroot.collapsed.nh \ + --vcf <(gunzip -c gisaid-$releaseLabel.vcf.gz) \ +| tail -n+2 \ +| sed -re 's/^[A-Z]([0-9]+)[A-Z,]+.*parsimony_score=([0-9]+).*/\1\t\2/;' \ +| tawk '{print "NC_045512v2", $1-1, $1, $2;}' \ +| sort -k2n,2n \ + > gisaid-$releaseLabel.parsimony.bg +bedGraphToBigWig gisaid-$releaseLabel.parsimony.bg /hive/data/genomes/wuhCor1/chrom.sizes \ + gisaid-$releaseLabel.parsimony.bw + +# Full find_parsimonious_assignments output on collapsed tree for collaborators +time $find_parsimonious_assignments --tree ft_SH.reroot.collapsed.nh \ + --vcf gisaid-$releaseLabel.unfiltered.masked.vcf.gz \ + > find_parsimonious_assignments.$releaseLabel.collapsed.out + # Now filter by frequency for faster track display: alt al freq >= 0.001 sampleCount=$(wc -l < tree.renamedPruned.ids.sorted) minAc001=$(( (($sampleCount + 999) / 1000) )) time vcfFilter -minAc=$minAc001 -rename gisaid-$releaseLabel.vcf.gz \ > gisaid-$releaseLabel.minAf.001.vcf ls -l gisaid-$releaseLabel.minAf.001.vcf wc -l gisaid-$releaseLabel.minAf.001.vcf bgzip -f gisaid-$releaseLabel.minAf.001.vcf tabix -p vcf gisaid-$releaseLabel.minAf.001.vcf.gz # Alt al freq >= 0.01 minAc01=$(( (($sampleCount + 99) / 100) )) time vcfFilter -minAc=$minAc01 -rename gisaid-$releaseLabel.minAf.001.vcf.gz \ > gisaid-$releaseLabel.minAf.01.vcf ls -l gisaid-$releaseLabel.minAf.01.vcf wc -l gisaid-$releaseLabel.minAf.01.vcf bgzip -f gisaid-$releaseLabel.minAf.01.vcf tabix -p vcf gisaid-$releaseLabel.minAf.01.vcf.gz # Make sample color files #*** TODO: Nextstrain Clade is now in field 18! Add that as an option! cut -f3,18-20 $metadata \ | sort > metadata.epiToLineageAndClades # Join on EPI ID to associate tree sample names with lineages. join -t$'\t' tree.renaming metadata.epiToLineageAndClades -o 1.2,2.2,2.3,2.4 > sampleToLineage $scriptDir/cladeLineageColors.pl sampleToLineage gzip -f *Colors